Bioengineering a novel therapeutic protein complex to minimize the effects of medical device induced hemolysis

生物工程新型治疗性蛋白质复合物可最大程度地减少医疗设备引起的溶血的影响

• 批准号: 10542403
• 负责人: Paul Werner Buehler
• 金额: $ 65.37万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10542403
• 关键词: Acute Renal Failure with Renal Papillary Necrosis; Affinity; Animal Model; Animals; Attenuated; Binding; Binding Proteins; Binding Sites; Biological Assay; Biological Markers; Biomedical Engineering; Brain; Cardiac; Cardiac Output; Cardiac Surgery procedures; Cardiopulmonary Bypass; Cells; Cessation of life; Chemicals; Circulation; Clinical; Complex; Complication; Cytolysis; Data; Dimerization; Dose; Drug Metabolic Detoxication; Drug or chemical Tissue Distribution; Effectiveness; Engineering; Erythrocytes; Extracorporeal Membrane Oxygenation; Fractionation; Functional disorder; Goals; Hamsters; Haptoglobins; Health Care Costs; Heart; Heme; Hemoglobin; Hemoglobin concentration result; Hemolysis; Hemolysis Induction; Hemopexin; Hospitalization; Hour; Human; In Vitro; Individual; Injury to Kidney; Intestines; Iron; Kidney; Kinetics; Length of Stay; Ligands; Lung; Macrophage; Medical; Medical Device; Methods; Microvascular Permeability; Modeling; Monitor; Morbidity - disease rate; Operative Surgical Procedures; Organ; Outcome; Paste substance; Patients; Plasma; Procedures; Process; Proteins; Proteolysis; Public Health; Pump; Qualifying; Rattus; Reaction; Renal function; Risk; Sampling; Secure; Self-Help Devices; Severities; Supportive care; System; Testing; Therapeutic; Therapeutic Intervention; Therapeutic Uses; Toxic effect; Toxin; Transferrin; Urine; Ventricular; apohemoglobin; blood pump; cell injury; clinically relevant; cost; cost estimate; design; dimer; experience; heart function; hemodynamics; improved; in vivo; individual patient; instrument; large scale production; lung injury; monocyte; mortality; mortality risk; novel; novel therapeutics; organ injury; prevent; prospective; protein complex; proteostasis; pulmonary function; response; urinary

PROJECT SUMMARY Cost estimates for CNS, pulmonary, cardiac, and renal complications following medical assist devices requiring blood pumps, for example on pump cardiopulmonary bypass (CPB) is estimated at $80 million per individual states in the US over a ten-year period. Because of the need for extended CPB (> 4hours) use in complex surgeries, hemolysis, and the subsequent release of hemoglobin (Hb) into the circulation – which is known to be multi-organ toxic and vasoactive – is a relevant contributor to more intensive management of patients. To date, no effective strategy to remove toxic by-products of Hb are available. The burden caused by kidney injury alone accounts for approximately $9 billion/year, 300,000 deaths/year and an average increase in hospital stay of 3.5 days/patient This project focuses on understanding the contribution of hemolysis, Hb and heme associated with secondary end organ injury following extended on pump cardiopulmonary bypass (typically lasting up to 4 hours). To understand medical device related red blood cell lysis in this setting, we propose a strategy that is focused on sequentially understanding the contributions of Hb and heme and a therapeutic strategy to attenuate the end organ pathophysiology of these toxins. We will prospectively determine the levels of Hb, heme and iron as well as the concentrations of each toxin in plasma and urine of cardiac surgery patients. Simultaneously we will determine the concentrations of Hb, heme and iron binding and clearance proteins, haptoglobin, hemopexin and transferrin, respectively. Further we will determine plasma and urinary markers of end organ injury. We will use this data to construct a biokinetic model that determines the limits of Hb toxin concentrations that associate with end organ injury markers. The goal of this effort will be to define the need and timing for therapeutic interventions. To this end we have bioengineered a novel Hb, heme and protein scavenger based on our extensive experience with studying Hb toxicity. The novel protein construct is prepared by generating apo-Hb in multi-step process, while simultaneously isolating haptoglobin from Cohn fractionation paste IV. Finally, the two proteins are complexed and further purified to generate the apo-Hb-haptoglobin complex. This novel protein construct binds heme in high heme exposure states, secures iron in the heme ligand and safely clears the complex to monocytes and macrophages. Alternatively, in high Hb exposures apo-Hb exchanges binding sites on haptoglobin clearing Hb dimers to monocytes and macrophages, while released apo-Hb dimers are degraded by proteolysis and harmlessly cleared from circulation. To test the effect of the novel complex we have planed a range of proof-of-concept studies in animal models of on pump CPB to define the ability of apo- Hb-haptoglobin dosing to prevent end organ injury.

项目摘要 医疗辅助设备后，CNS，肺，心脏和肾脏并发症的成本估算值 需要血液泵，例如，在泵的心肺旁路（CPB）上，估计为8000万美元 在十年内，美国的个别州。因为需要扩展CPB（> 4小时） 复杂的手术，溶血以及随后的血红蛋白（HB）释放到循环中 - 众所周知有毒和血管活性 - 是对更密集管理的相关贡献者 患者。迄今为止，尚无有效的策略去除HB的有毒副产品。烧伤造成了 仅通过肾脏伤害，每年约为90亿美元，每年30万人死亡和平均 住院时间增加3.5天/患者该项目的重点是了解 溶血，HB和血红素与次级末端器官损伤相关 心肺旁路（通常持续长达4个小时）。了解医疗装置相关的红细胞 在这种情况下，我们提出了一种策略，该策略侧重于依次了解 HB和Heme以及一种减弱这些毒素末端器官病理生理学的治疗策略。我们将 前瞻性确定Hb，血红素和铁的水平以及每种毒素的浓度 心脏手术患者的血浆和尿液。同时我们将确定 HB，血红素和铁结合和清除蛋白，触觉球蛋白，血红素和转移蛋白分别。 此外，我们将确定最终器官损伤的血浆和尿标志物。我们将使用这些数据来构建 一个确定与最终器官损伤相关的HB毒素浓度极限的生物动力学模型 标记。这项工作的目的是定义治疗干预的需求和时间。对此 最后，我们根据我们的丰富经验，为生物工程设计了一种新颖的HB，血红素和蛋白质清道夫 研究HB毒性。新颖的蛋白质构建体是通过在多步骤过程中生成apo-hb来制备的， 同时从科恩分级糊剂中分离出触蛋白IV。最后，两个蛋白质是 复合并进一步纯化以生成Apo-Hb-Haptoglobin复合物。这个新颖的蛋白质构建 在高血红素暴露状态下结合血红素，在血红素配体中固定铁并安全地清除复合物 单核细胞和巨噬细胞。另外，在高HB中，APO-HB交换位点上的位点 将HB二聚体清除为单核细胞和巨噬细胞，而释放的Apo-Hb二聚体降解 通过蛋白水解和无害的循环清除。为了测试新颖复合物的效果，我们有 计划在On Pump CPB的动物模型中进行一系列概念验证研究，以定义apo-的能力 HB-热蛋白给药以防止最终器官损伤。